小柯机器人

德国马克斯普朗克生物化学研究所Ralf Jungmann团队近期取得重要工作进展，他们研究提出了对单个蛋白质水平上绝对标记效率的量化的方法。相关研究成果2024年4月24日在线发表于《自然—方法学》杂志上。

据介绍，最先进的超分辨率显微镜使研究人员能够在空间上分辨密集簇中的单个蛋白质。然而，蛋白质组织和化学计量的精确量化需要一种通用的方法来评估绝对的粘合剂标记效率。

研究人员介绍了一种普遍适用的方法，该方法使用与感兴趣的靶蛋白融合的参考标签。通过将高亲和力的结合物（如抗体或纳米体）连接到参考标签和靶蛋白上，然后使用DNA条形码序列超分辨率成像，研究人员可以将参考标签的位置与靶分子结合物相关联。这种方法有助于在单个蛋白质水平上精确量化标记效率。

附：英文原文

Title: Quantification of absolute labeling efficiency at the single-protein level

Author: Hellmeier, Joschka, Strauss, Sebastian, Xu, Shuhan, Masullo, Luciano A., Unterauer, Eduard M., Kowalewski, Rafal, Jungmann, Ralf

Issue&Volume: 2024-04-24

Abstract: State-of-the-art super-resolution microscopy allows researchers to spatially resolve single proteins in dense clusters. However, accurate quantification of protein organization and stoichiometries requires a general method to evaluate absolute binder labeling efficiency, which is currently unavailable. Here we introduce a universally applicable approach that uses a reference tag fused to a target protein of interest. By attaching high-affinity binders, such as antibodies or nanobodies, to both the reference tag and the target protein, and then employing DNA-barcoded sequential super-resolution imaging, we can correlate the location of the reference tag with the target molecule binder. This approach facilitates the precise quantification of labeling efficiency at the single-protein level. Super-resolution imaging of reference and target structures enables precise determination of the labeling efficiency of high-affinity binding proteins in cells for improved quantitative assessment of protein organization at the single-molecule level.

DOI: 10.1038/s41592-024-02242-5

Source: https://www.nature.com/articles/s41592-024-02242-5

Nature Methods：《自然—方法学》，创刊于2004年。隶属于施普林格·自然出版集团，最新IF：47.99
官方网址：https://www.nature.com/nmeth/
投稿链接：https://mts-nmeth.nature.com/cgi-bin/main.plex